The role of WC1(+) gamma delta T-cells in the delayed-type hypersensitivity (DTH) skin-test reaction of Mycobacterium bovis-infected cattle

Delayed-type hypersensitivity (DTH) skin-testing with mycobacterial antigens is often used as a means of identifying Mycobacterium bovis-infected cattle. Better understanding of the cellular basis underlying the DTH reaction is required if diagnostic methods are to be improved upon. Previous studies have shown that gamma delta T-cells, particularly those bearing the WC1 molecule, are present at an early stage of developing DTH responses and that such cells may modulate the developing immune response immediately following M. bovis-infection. However, their role, if any, in the DTH response remains unclear. In the present study we have used an in vivo model to deplete WC1(+) gamma delta T-cells, from cattle with established M. bovis-infection, prior to skin-testing. Results indicate that, although WC1(+) gamma delta T-cells do infiltrate the skin-test site in normal calves, they do not appear to be essential for the development of DTH skin swelling, as indicated by effective development of skin responses in calves depleted of circulating WC1(+) gamma delta T-cells.     

Primary meningeal T-cell lymphoma in a harbor seal (Phoca vitulina).

A 15-year-old female harbor seal (Phoca vitulina) was referred to the Nantes Veterinary School, Nantes, France, with a clinical history of anorexia, seizures, and left hemiplegia. Cerebrospinal fluid analysis revealed large numbers of neoplastic lymphoid cells. After injection of a contrast agent into the cerebrospinal space, radiographs demonstrated an asymmetry of the right lateral ventricle. Necropsy examination revealed a marked edema of the right frontal lobe, extending to the basal nuclei and thalamus in the brain. Histological examination of the brain revealed leptomeningeal lymphoma. Immunohistochemical labeling demonstrated that the neoplasm was of T-cell origin. No significant macroscopic or microscopic lesions were observed in the other organs examined, including lymphoid organs. This is the first report of primary leptomeningeal lymphoma in a harbor seal.     

IFN gamma and IL-4 production by CD4, CD8 and WC1 gamma delta TCR(+) T cells from cattle lymph nodes and blood

The synthesis of IFN gamma and IL-4 by CD4, CD8 and WC1 gamma delta TCR(+) T cell sub-populations, and T cells stained with activation/memory-sub-set markers has been examined by flow cytometric analysis. Cells from blood, prescapular, bronchial and mesenteric lymph nodes and Peyer's patches were incubated with phorbol 12-myristate 13-acetate (PMA), ionomycin and brefeldin-A before staining. Lymphocytes that stained for cytoplasmic IFN gamma were evident within the CD4 and CD8 populations from all tissues and also in the WC1 population from lymph nodes. IL-4 producing cells were primarily evident within the CD4 population. IFN gamma synthesis was evident within both CD45RO(+) and CD45RB(+) populations, but IL-4 synthesis was predominantly by cells that were CD45RO(+)/CD45RB(-). Expression of CD62L is not related to functional memory in CD4(+) T cells from cattle and CD62L(+) cells, particularly from the lymph nodes draining the skin and the lungs, stained with mAb to IFN gamma and IL-4. The findings indicate that at least for CD4(+) T cells, where CD45 isoform expression is related to functional memory, these two cytokines are produced predominantly by cells with a memory phenotype. The observation that some WC1(+) cells produce IFN gamma implies the presence of distinct sub-sets of this gamma delta TCR(+) population cattle and suggests a functional role.     

Gamma/delta T-cell lymphoma in a dog

A 15-month-old dachshund was presented for examination because of a cough. Thoracic radiographs revealed the presence of a mass in the upper mediastinum. A diagnosis of γδ T-cell lymphoma was made by biopsy and flow cytometry analysis. The dog was treated with chemotherapy and remains asymptomatic after 24 months.     

Canine T-cell lymphomas: a morphological,immunological, and clinical study of 46 new cases

The aim of this study is to report 46 new cases of canine T-cell lymphomas among a series of 140 lymphomas studied by immunophenotyping (incidence 32.8%). According to the updated Kiel classification adapted to the canine species, 13 were classified as low-grade and 33 as high-grade lymphomas. Among the low-grade lymphomas, five were small clear-cell lymphomas, three were pleomorphic small-cell lymphomas, and five mycosis fungoides. Among the high-grade cases, there were 11 pleomorphic mixed-, small-, and large-cell lymphomas, 6 pleomorphic large-cell lymphomas, 11 lymphoblastic lymphomas, and 5 unclassifiable high-grade plasmacytoid lymphomas. The cytohistologic features were highly suggestive of a T-cell phenotype on the basis of cell morphology (irregular nuclei and clear cytoplasms) (30/46 cases), a T-cell zone pattern, and the presence of hyperplastic postcapillary venules (22/46 cases). All 46 cases were CD3+ CD79a-, and among 34 cases investigated for CD4 and CD8 expression, 13 were CD4+CD8-, 13 were CD8+CD4-, and 8 were CD4CD8 double positive or double negative. The pleomorphic mixed lymphomas were mainly CD4+CD8- (6/7) and the lymphoblastic lymphomas were double positive or double negative (6/8). The main clinical, hematologic, and biochemical features were generalized (28/46) or regional lymphadenopathy (16/46), hepatosplenomegaly (15/46), extranodal involvement (11/46), mediastinal mass (9/46), and leukemia (8/46), which were mainly present in cases of lymphoblastic lymphomas and hypercalcemia (16/46).     

SIV-associated lymphomas in rhesus monkeys (Macaca mulatta) in comparison with HIV-associated lymphomas

A retrospective study was performed to characterize malignant lymphomas of 16 Simian immunodeficiency virus (SIV)-infected rhesus monkeys (Macaca mulatta), 2-9 years of age, on the basis of clinical data, histologic and immunophenotypic results, and cell death indices compiled with the TdT-mediated X-duTP nick end labeling method. We particularly focused on providing immunohistochemical evidence of expression products of EBNA2, Bc12, c-Myc, P21, P53, and Bc16. Results were compared with data from the literature on human HIV-associated lymphomas. According to the updated Kiel classification, the lymphomas were classified as 11 centroblastic lymphomas, three immunoblastic lymphomas, one Burkitt-like lymphoma, and one immunocytoma. Using antibodies to CD20, the B-cell origin of tumor cells was demonstrated. SIV antigen was not demonstrated in the tumor cells. Infection with rhesus lymphocryptovirus was present in 94% of the monkeys. Lymphomas revealed expression of Bc12 in 15/16 (94%), c-Myc in 14/16 (88%), P21 in 10/ 16 (63%), P53 in 12/16 (75%), and Bc16 in 1/16 (6%) monkeys. This study provided evidence that the expression of these gene products, which are thought to play an important role in cell proliferation and apoptosis in HIV- and non-HIV-associated lymphomas, are also involved in the pathogenesis of lymphomas in SIV-infected rhesus monkeys. A tentative relationship between the described gene products and the cell death indices was established for the expression of Bc12. The present primate model represents a suitable animal model for studying the pathogenesis of AIDS-associated lymphomas.     

Characterization of C-, J- and V-region-genes of the feline T-cell receptor gamma

Lymphomas and leukemias are important neoplasias of domestic cats and human beings. In some cases it can be difficult to differentiate these tumors from reactive lymphatic hyperplasia. To overcome this problem, the diagnosis of lymphomas and leukemias in man is often supported by molecular techniques. To be able to establish such a technique in the cat we had to sequence the genes coding for the antigen receptors. As primary target in this study we choose the T-cell receptor gamma. Using 5'-and 3'-RACE techniques we were able to clone and sequence four different V-region genes, which can be clustered into two subgroups as well as six variants of the C-region gene. Additionally, we found eight J-region genes which can be classified into three subgroups. One of the V-region genes, six of the J-region genes and all C-region genes had not been described previously. All together we analysed 112 clones containing V- and J-region genes and 31 clones containing C-region genes. Sixty-six of these clones were full length containing the L-region as well as the 5'-UTR of the feline T-cell receptor gamma. The sequences of the V-region- and J-region-genes show sufficiently homologous areas that can be used to establish a small number of consensus-primers to be applied in molecular diagnosis of feline lymphomas and leukemias.     

Association between postpartum pyrexia and uterine bacterial infection in dairy cattle

The temperature of 90 dairy cattle was recorded for the first 10 days after parturition and the animals were categorised as either normal (< 39.7 degreesC) or pyrexic. Swabs were collected from the uterine lumen seven, 14, 21 and 28 days after parturition for aerobic and anaerobic culture; bacteria were identified and their growth was scored semiquantitatively. Blood samples were collected three times a week for the estimation of the concentrations of acute phase proteins. The cows' temperatures were often above the accepted normal range, but it was not a good indicator of the number of bacteria in the uterus. However, pyrexia was correlated with the presence of specific uterine pathogens (P < 0.05) and in particular with Prevotella species (P < 0.01). The pyrexic animals had a higher plasma concentration of the acute phase protein (alpha1-acid glycoprotein (P < 0.05). Although pyrexia is an indicator of postpartum inflammation, additional clinical signs are necessary to identify uterine bacterial infection.     

Disposition of ampicillin trihydrate in plasma,uterine tissue,lochial fluid,and milk of postpartum dairy cattle

The objective of this study was to determine the disposition of ampicillin in plasma, uterine tissue, lochial fluid, and milk of postpartum dairy cattle. Ampicillin trihydrate was administered by intramuscular (i.m.) injection at a dose of 11 mg/kg of body weight every 24 h (n = 6, total of 3 doses) or every 12 h (n = 6, total of 5 doses) for 3 days. Concentrations of ampicillin were measured in plasma, uterine tissue, lochial fluid, and milk using HPLC with ultraviolet absorption. Quantifiable ampicillin concentrations were found in plasma, milk, and lochial fluid of all cattle within 30 min, 4 h, and 4 h of administration of ampicillin trihydrate, respectively. There was no significant effect of dosing interval (every 12 vs. every 24 h) and no significant interactions between dosing interval and sampling site on the pharmacokinetic variable measured or calculated. Median peak ampicillin concentration at steady‐state was significantly higher in lochial fluid (5.27 μg/mL after q 24 h dosing) than other body fluids or tissues and significantly higher in plasma (3.11 μg/mL) compared to milk (0.49 μg/mL) or endometrial tissue (1.55 μg/mL). Ampicillin trihydrate administered once daily by the i.m. route at the label dose of 11 mg/kg of body weight achieves therapeutic concentrations in the milk, lochial fluid, and endometrial tissue of healthy postpartum dairy cattle.     

Serum gamma glutamyl transpeptidase activity in cattle with induced fascioliasis

Six-month-old calves were inoculated with 200 metacercariae of Fasciola hepatica over a 72 day period. Their aspartic transaminase, alkaline phosphatase, arginase and gamma glutamyl transpeptidase (GGT) values were measured at two-week intervals. Infections with a final fluke count of 19 to 87 flukes did not result in an increase of serum alkaline phosphatase or arginase compared with those in control calves. Serum aspartic transaminase values were increased significantly (P less than 0.025) above those of controls but were not increased over reference values. Seemingly, measurement of these enzymes would not be useful in assessing the degree of fluke infestation. In contrast, serum values of GGT increased 56 days after preliminary inoculation and remained high for at least 83 days after infection. The initial rise coincided with the penetration of bile ducts by migrating flukes. This relationship may be useful in further studies of fluke-induced biliary damage.     

Contributions of ovum cytoplasm, uterine environment and postnatal environment to maternal effects in beef cattle

In the first part of the experiment, embryos of Brown Swiss-Hereford (BS-H) reciprocal crosses were transferred to BS and H recipient cows and embryos of Red Poll-Angus (RP-A) reciprocal crosses were transferred to RP and A recipient cows. Calves resulting from embryo transfers (ET) were weaned early (3 to 5 d). Males were castrated and their growth, feed efficiency and carcass composition were recorded. In the second part of the experiment, matings to produce reciprocal crosses were made by natural service (NS) between BS and H and between RP and A. Some of the offspring were weaned at 3 to 5 d; the remainder nursed their dams to an age of 150 to 180 d. Growth after 200 d, feed efficiency and carcass data were recorded on steers. Females produced in both parts of the experiment were reared for breeding, and post-200-d growth and age at puberty were recorded. Neither breed of recipient (uterine influence) nor breed of donor (cytoplasmic influence) had important effects on growth or carcass traits. Large differences were observed for most growth and carcass traits between offspring that were weaned early and those that nursed their dams. Effects of early weaning were greater in progeny of Brown Swiss and Red Poll dams than in progeny of Angus and Hereford dams. Prenatal maternal influences (e.g., ovum cytoplasm and uterine influence) are relatively small in comparison with postnatal maternal influences in contributing to maternal effects in beef cattle.     

Bovine lineage-specific gamma delta (gammadelta) T cell GD3.5 antibody cross-reacts with cell surface antigens on ovine and caprine lymphocytes

A specifically defined function for gammadelta T cells is still a topic of intense debate. Thus, there is great importance in the identification of lineage-specific markers that characterize these cells. Early studies describe a family of lineage-specific cell-surface molecules on ruminant gammadelta T cells called WC1. More recently, a 220-240 kDa lineage-specific bovine gammadelta T cell surface marker, GD3.5Ag, was reported. Here, we used anti-bovine GD3.5Ab to study its cross-reactivity with caprine and ovine lymphocytes. Flow cytometric analysis demonstrated that GD3.5Ab binds ovine and caprine lymphocytes. In addition, immunoprecipitation experiments with GD3.5Ab demonstrate an ovine and caprine cross-reactive antigen that is similar in size to the bovine antigen. These results suggest that GD3.5Ag is well conserved among ruminant species and GD3.5Ag may play an important role in gammadelta T cell biology.     

Cytopathology of canine lymphomas (100 cases)

Malignant lymphoma is one of the most common malignant tumours occurring in dogs. Fine needle aspiration biopsy (FNAB) is an excellent, specific diagnostic procedure used to assess pathological processes in lymph nodes. The aim of the present study was to conduct a cytopathological analysis of lymphoma in dogs and to analyse some epidemic aspects of occurrence of lymphoma in 100 dogs using Giemsa stained slides. Samples were obtained by fine needle aspiration biopsy, fine needle non-aspiration biopsy, lymph node impression smears and by examination of body cavity effusions. The determination of the type and subtype of tumour was made on the basis of the updated Kiel classification adopted for dogs. Based on cytopathological analysis, the lymphoma was diagnosed in 100 dogs: 44 were female and 56 male. The animals' age ranged from 1.5 year to 15 years (median: 7.5 years), the animals were of different breeds (72% of all dogs belonged to 28 different breeds) and crossbreeds (28%). In 29% of dogs the regional or general lymphadenomegaly was the only clinical sign observed, in remaining cases (71%) at least one abnormality connected to lymphoma was found. Among all diagnosed lymphomas, high-grade lymphomas were more prevalent (86% of all cases) than low-grade lymphomas (14% of all cases). The possibility of boxers having a predisposition to T cell lymphoma development could be also suspected.     

Experimental cross-infections of Haemonchus placei (Place, 1893) in sheep and cattle

To examine effects on biology and morphology, Haemonchus placei infections were propagated in cattle or sheep and infective larvae were introduced into the same or opposite host. Ovine source larvae had a geometric mean (GM) prepatent period of 26.5 days regardless of host species, compared to 30.6 days for bovine source larvae regardless of host species. Similarly, ovine source H. placei had a higher GM percentage establishment versus bovine source larvae (9.6% versus 3.4%) regardless of host species. The patent daily egg count for ovine source versus bovine source H. placei was 109.1 versus 50.0 FEC regardless of host species. Mean spicule length for ovine source H. placei was 492.5 microm while bovine source measured 496.5 microm. Mean female tail length for ovine source H. placei was 586.5 and 589.5 microm for bovine source H. placei. Using synlophe morphology, all worms that were measured were identified as H. placei. Sixty-three percent of females examined for vulvar flap morphology had knob-like vulvar flaps while the remaining 37% had linguiform vulvar flaps.